Allele-specific amplification for preimplantation genetic diagnosis (PGD) of spinal muscular atrophy

We have developed a new allele-specific amplification method for the preimplantation genetic diagnosis (PGD) of spinal muscular atrophy (SMA; Werdnig-Hoffmann disease) from a single cell. This method is based on the detection of the deletion of exon 7 of the telomeric copy of the survival motor neurone (SMN^t) gene. An oligonucleotide was designed to be specific to the SMN^t nucleotidic sequence with exonic mismatch G (for SMN^t)→A (for SMN^c) at its 3′ end. This test produces reliable PCR products in 95% of single lymphoblasts (85/88) tested as well as in 16/16 blastomeres from normal controls. Specificity analysis showed that we were able to detect homozygous deletion of the SMN^t gene in 99% of single lymphoblasts (103/104) from a SMA patient. No contamination was detected in 68 blanks tested. Multiple cell and DNA dilution analysis revealed that the test is accurate and specific up to 100 pg DNA and should thus also be suitable for PGD at the blastocyst stage. This rapid procedure requires a single round of fluorescent PCR and no restriction digestion, while previously described single cell methods include nested PCR followed by restriction enzyme digestion. Two PGD cycles for SMA using this procedure were performed in our centre. Copyright © 2001 John Wiley & Sons, Ltd.     

Prenatal diagnosis of exophytic nevus sebaceous of the scalp

Nevus sebaceous is a complex hamartoma most commonly found on the scalp, face, and neck and is often present at birth, although some may be diagnosed later in infancy. We report the first prenatal diagnosis of isolated nevus sebaceous that presented at 19 weeks' gestation as a large and exophytic tumor of the scalp. This case emphasizes the crucial role of ultrasound examination performed with high-frequency probes, which revealed associated diffuse lesions of the face. Identification of such a facial involvement could have a dramatic prognostic impact. © 2013 John Wiley & Sons, Ltd.     

Prevalence of Human Noroviruses in Frozen Marketed Shellfish,Red Fruits and Fresh Vegetables

Noroviruses (NoVs), currently recognised as the most common human food-borne pathogens, are ubiquitous in the environment and can be transmitted to humans through multiple foodstuffs. In this study, we evaluated the prevalence of human NoV genogroups I (GI) and II (GII) in 493 food samples including soft red fruits (n = 200), salad vegetables (n = 210) and bivalve mollusc shellfish (n = 83), using the Bovine Enterovirus type 1 as process extraction control for the first time. Viral extractions were performed by elution concentration and genome detection by TaqMan Real-Time RT-PCR (RT-qPCR). Experimental contamination using hepatitis A virus (HAV) was used to determine the limit of detection (LOD) of the extraction methods. Positive detections were obtained from 2 g of digestive tissues of oysters or mussels kept for 16 h in seawater containing 2.0–2.7 log₁₀ plaque-forming units (PFU)/L of HAV. For lettuces and raspberries, the LOD was, respectively, estimated at 2.2 and 2.9 log₁₀ PFU per 25 g. Of the molluscs tested, 8.4 and 14.4 % were, respectively, positive for the presence of GI NoV and GII NoV RNA. Prevalence in GI NoVs varied from 11.9 % for the salad vegetables samples to 15.5 % for the red soft fruits. Only 0.5 % of the salad and red soft fruits samples were positive for GII NoVs. These results highlight the high occurrence of human NoVs in foodstuffs that can be eaten raw or after a moderate technological processing or treatment. The determination of the risk of infection associated with an RT-qPCR positive sample remains an important challenge for the future.     

Potential of Pulsed Light to Inactivate Bacteriophage MS2 in Simple Liquid Medium and on Complex Foodstuffs

The virucidal efficacy of a pulsed light treatment applied to viral suspensions, glass beads and herb powders was studied for the F-RNA bacteriophage MS2. The experimental results obtained demonstrated the high potential of this technology to efficiently decontaminate simple matrices but underlined the complexity of application to complex food matrices.     

Analysis of PFAAs in American alligators part 2: Potential dietary exposure of South Carolina hunters from recreationally harvested alligator meat

Exposure to perfluorinated alkyl acids (PFAAs) has been linked to many harmful health effects including reproductive disorders, developmental delays, and altered liver and kidney function. Most human exposure to environmental contaminants, including PFAAs, occurs through consumption of contaminated food or drinking water. This study uses PFAA data from meat samples collected from recreationally harvested American alligators (Alligator mississippiensis) in South Carolina to assess potential dietary exposure of hunters and their families to PFAAs. Consumption patterns were investigated using intercept surveys of 23 hunters at a wild game meat processor. An exposure scenario using the average consumption frequency, portion size, and median perfluorooctane sulfonic acid (PFOS) concentration in alligator meat from all hunt units found the daily dietary exposure to be 2.11 ng/kg body weight per day for an adult human. Dietary PFOS exposure scenarios based on location of harvest suggested the highest daily exposure occurs with alligator meat from the Middle Coastal hunt unit in South Carolina. Although no samples were found to exceed the recommended threshold for no consumption of PFOS found in Minnesota state guidelines, exposure to a mixture of PFAAs found in alligator meat and site-specific exposures based on harvest location should be considered in determining an appropriate guideline for vulnerable populations potentially exposed to PFAAs through consumption of wild alligator meat.     

Modeling fecundity in birds: Conceptual overview, current models, and considerations for future developments

Fecundity is fundamental to the fitness, population dynamics, conservation, and management of birds. For all the efforts made to measure fecundity or its surrogates over the past century of avian research, it is still mismeasured, misrepresented, and misunderstood. Fundamentally, these problems arise because of partial observability of underlying processes such as renesting, multiple brooding, and temporary emigration. Over the last several decades, various analytical approaches have been developed to estimate fecundity from incomplete and biased data. These, include scalar arithmetic formulae, partial differential equations, individual-based simulations, and Markov chain methodology. In this paper, we: (1) identify component processes of avian reproduction; (2) review existing methods for modeling fecundity; (3) place these diverse models under a common conceptual framework; (4) describe the parameterization, validation, and limitations of such models; and (5) point out future considerations and challenges in the application of fecundity models. We hope this synthesis of existing literature will help direct researchers toward the most appropriate methods to assess avian reproductive success for answering questions in evolutionary ecology, natural history, population dynamics, reproductive toxicology, and management.